Figure 4. Analyses of astrocytes subjected to pressure for iso[4]LGE₂–protein adduct formation. A: ELISA analysis was performed with antibodies to iso[4]LGE₂–protein adduct and 10 µg of the control and pressure-treated astrocyte lysate (described in greater detail in Methods). Following pressure treatment, the culture medium was immediately replaced with fresh medium, and the cells were incubated at 37 °C for 16 h at atmospheric pressure. Immunocytochemical analysis with iso[4]LGE₂–protein adduct antibody was carried out for control cells (B) and for astrocytes (C) that were subjected to 3 h of pressure at 100 mmHg and then subsequently subjected to a recovery period of 16 h.